Mda mb 231 luc

A549-luc, BXPC-3-luc, HT-29-luc, Hela-luc, MDA-MB231-luc, NCIH460-luc, PC-3M-luc, and LL/2-luc-M38 cell lines were all purchased from Caliper Life Sciences. U87MG-luc cells were generated by our group previously [35 ]. The primary culture was derived from freshly isolated tumor cell from patients of two primary lung cancer or two primary breast cancer individually, in Beijing Shijitan Hospital of Capital Medical University. The consent was obtained from the patients before sample collection. The study complied with the Declaration of Helsinki and was approved by the Biomedical Research Ethics Committee of Beijing Shijitan Hospital of Capital Medical University. Briefly, tumors were placed into RPMI1640 medium with antibiotics and anti-fungal immediately following resection. Specimens were washed with cold PBS repeatedly to remove blood clots and tissue debris, and then minced into 0.5- to 1.0-mm pieces. The tissue pieces were digested with collagenase IV, Dnase I and hyaluronidase. Then cells were washed and passed through a 70 μm cell strainer to obtain single-cell population. All agents for cell culture were from Gibco Company (Gaithersburg, MD, USA). Beige-SCID Mice (8 to 10 weeks) were purchased from Vital River Laboratories (VRL, Beijing, China).

Human cancer cell lines, A2058, AsPC-1, SKNSH, MiaPaCa-2, cfPac-1, NCI-H460, H1915, H1299, U87MG and U373 and the normal pancreatic cell line (HPDE) were obtained from ATCC (Manassas, VA, USA). MDA-MB-231-luc- were obtained from Caliper Life Sciences (Mountain View, CA, USA). HUVEC were from Lonza (Basel, Switzerland). HPDE, A2058, AsPC-1, MiaPaCa-2, cfPac-1, U87MG, Gli36 and U373 were cultured in DMEM (Fisher Scientific, Pittsburgh, PA, USA). H460, H1299 and H1915 were cultured in RPMI 1640 (Fisher Scientific). MDA-MB-231- Luc and SKNSH were cultured in AMEM (Invitrogen, Carlsbad, CA, USA). The above cell lines except HUVEC, were cultured in their respective media supplemented with 10% FBS and 1% Penicillin/Streptomycin. HUVEC were grown in EGM-2 media (Lonza).

HEK 293T human embryonic kidney cells, MDA-MB231 human breast tumor cells, MCF-7 human breast tumor cells, 4T1 mouse breast tumor cells, and Panc-1 human pancreatic tumor cells were purchased from American Type Culture Collection (ATCC, Rockville, MD). MDA-MB231-luc and 4T1-luc that express luciferase were obtained from Perkin Elmer (Waltham, MA). All cells were grown in Dulbecco's modified Eagle's medium (DMEM, HyClone, South Logan, UT) except MCF-7 cells, which were grown in Roswell park memorial institute medium (RPMI, HyClone). All media were supplemented with 10% fetal bovine serum (FBS, ThermoFisher Scientific, Waltham, MA). Cells were maintained at 37 °C in a humidified atmosphere containing 5% CO₂.

PF-03084014 and sunitinib were synthesized by Pfizer chemists. Unless otherwise noted, all chemicals were purchased from Sigma-Aldrich (St. Louis, MO). MDA-MB-231Luc and AngioSense 680 EX was purchased from PerkinElmer (Waltham, MA). The antibodies for IHC analyses were anti-BrdU (BD Pharmingen, San Diego, CA), anti-HIF1α (R&D Systems, Minneapolis, MN), anti-phospho-H2AX, anti-HES1, and anti-VEGFR2 (Cell Signaling Technology, Danvers, MA).